CN114761803A

CN114761803A - Methods of diagnosing cancer using anti-BAG 2 antibodies

Info

Publication number: CN114761803A
Application number: CN202080028383.7A
Authority: CN
Inventors: 金圣镇; 梁境珉; J·郑; S·G·安; S·H·朴; D·W·康
Original assignee: Sungkyunkwan University School Industry Cooperation; Industry Academic Cooperation Foundation of Yonsei University; Medpacto Inc
Current assignee: Sungkyunkwan University School Industry Cooperation; Industry Academic Cooperation Foundation of Yonsei University; Medpacto Inc
Priority date: 2019-02-12
Filing date: 2020-02-12
Publication date: 2022-07-15
Also published as: MX2021009690A; AU2020222326A1; WO2020165797A1; JP2022521706A; BR112021015940A2; SG11202108140SA; EP3938788A4; EP3938788A1; EP3924382A4; US20230183324A1; CN114269779A; JP2022525843A; WO2020165794A1; CA3129748A1; US20230194534A1; US12122822B2; EP3924382A1

Abstract

Disclosed herein is a composition for diagnosing cancer, comprising: an antibody or antigen-binding fragment thereof that specifically binds to a BAG2 polypeptide or fragment thereof.

Description

Methods of diagnosing cancer using anti-BAG 2 antibodies

Background

1. The technical field is as follows:

the present disclosure relates to compositions for diagnosing cancer comprising an antibody or antigen-binding fragment thereof that specifically binds to a BAG2 polypeptide or fragment thereof; and methods of providing information useful in diagnosing cancer.

2. General background and prior art:

the family of co-chaperone Bcl-2-associated immortal gene (BAG) proteins mediate a variety of physiological processes including intracellular protein folding, stress response, neuronal differentiation, apoptosis, and cell proliferation, and are functionally associated with a variety of synergistic proteins. BAG2, one of the BAG domain family members with anti-apoptotic activity, is a negative regulator of the C-terminus of the Hsc70 interacting protein (CHIP), a chaperonin-related ubiquitin ligase. The major role of BAG2 in the regulation of proteins by inhibition of CHIP activity is related to neurodegenerative and autosomal recessive diseases by stabilizing chaperonin-related proteins (e.g. PINK1 and CFTR). BAG2 is reported to have pro-apoptotic activity in a manner such that expression of BAG2 increases proteasome inhibitor-induced apoptosis, and BAG2 knockdown may partially inhibit apoptosis when thyroid cancer cells are exposed to proteasome inhibitor MG 132. It has also been reported that overexpression of BAG2 promotes stabilization of the potent tumor gene STK33 protein in various mutant K-Ras-induced tumors, thereby promoting tumor development. However, despite these findings, the role of BAG2 in cancer progression and metastasis is not clear.

Therefore, there is a need to develop compositions and methods for diagnosing cancer using antibodies that specifically bind to BAG2 polypeptide or fragments thereof.

Disclosure of Invention

These and other objects of the present invention will be more fully understood from the following description of the invention, the appended drawings and the appended claims.

One aspect provides a composition for diagnosing cancer comprising an antibody or antigen-binding fragment thereof that specifically binds to a BAG2 polypeptide or fragment thereof.

Another aspect provides a kit for diagnosing cancer, the kit comprising the composition.

Another aspect provides a method of providing information for diagnosing cancer.

Additional aspects will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the presented embodiments of the disclosure.

BAG2 polypeptides may be derived from a mammal. The mammal may be a human (homo sapiens), a mouse (mus musculus), a monkey, a cow or a horse. BAG2 may comprise the amino acid sequence of SEQ ID NO: 69. The amino acid sequence of SEQ ID NO 69 is the sequence corresponding to NCBI reference SEQ ID NO NM-004282.4. The BAG2 protein contains variants with bioequivalent activity to the amino acid sequence of SEQ ID NO:69, despite their amino acid sequence mismatch with the amino acid sequence of SEQ ID NO: 69. The BAG2 polypeptide may comprise an amino acid sequence having at least 60%, e.g., at least 70%, at least 80%, at least 90%, at least 95%, at least 99%, or 100% sequence identity to the sequence of SEQ ID No. 69. The BAG2 protein may be a polypeptide having a sequence identical to SEQ ID No. 69 except at least one amino acid residue, at least two amino acid residues, at least three amino acid residues, at least four amino acid residues, at least five amino acid residues, at least six amino acid residues, or at least seven amino acid residues. In the present specification, "polypeptide" may be used interchangeably with "protein".

Antibodies refer to specific immunoglobulins directed against an antigenic site. An antibody refers to a polypeptide or combination of polypeptides that specifically binds to BAG2 polypeptide or fragment thereof. The antibody includes polyclonal antibodies, monoclonal antibodies or recombinant antibodies such as ScFv fragments, diabodies, single-chain antibodies, etc., and includes all immunoglobulin antibodies. Antibodies may include antibodies having the complete form of two full-length light chains and two full-length heavy chains, and may also include functional fragments of antibody molecules thereof that retain antigen binding function by virtue of containing a specific antigen binding site (i.e., binding domain), despite the absence of the structure of the complete antibody having the complete form of two light chains and two heavy chains.

The antigen-binding fragment is a fragment of the entire structure of an immunoglobulin, and refers to a portion of the polypeptide that comprises a portion capable of binding to an antigen. For example, the antigen binding fragment can be an scFv, (scFv)2, Fv, Fab ', Fv F (ab')2, or a combination thereof.

There are five heavy chains, gamma, delta, alpha, mu and epsilon, and the heavy chains may determine the type of antibody. α and γ each comprise 450 amino acids, and μ and ε each comprise 550 amino acids. The heavy chain has two regions, a variable region and a constant region.

Two light chains, κ and λ, are present and may comprise from about 211 amino acids to about 217 amino acids. Light chains can have constant and variable regions.

The antibody or antigen-binding fragment thereof may comprise: a heavy chain variable region (which comprises complementarity determining region (VH-CDR)1 consisting of the amino acid sequence of SEQ ID NO:33, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:39, and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45), and a light chain variable region (which comprises complementarity determining region (VL-CDR)1 consisting of the amino acid sequence of SEQ ID NO:51, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:57, and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63);

a heavy chain variable region (which comprises VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:34, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:40 and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 46), and a light chain variable region (which comprises VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:52, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:58 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 64);

a heavy chain variable region (the heavy chain variable region comprising VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:35, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:41 and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 47), and a light chain variable region (the light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:53, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:59 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 65);

a heavy chain variable region (the heavy chain variable region comprising VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:36, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:42 and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 48), and a light chain variable region (the light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:54, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:60 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 66);

a heavy chain variable region (which comprises VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:37, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:43 and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 49), and a light chain variable region (which comprises VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:55, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:61 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 67); and

a heavy chain variable region (the heavy chain variable region comprising VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:38, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:44 and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 50), and a light chain variable region (the light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:56, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:62 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 68); or a combination thereof.

Xaa 6 and 7 in SEQ ID NO:39 can be glycine (Gly) or alanine (Ala). The second Xaa in SEQ ID NO 35 can be tyrosine (Tyr) or histidine (His). The 8 th Xaa in SEQ ID NO:41 can be serine (Ser) or threonine (Thr). The 12 th Xaa in SEQ ID NO:47 can be tyrosine (Tyr) or histidine (His). The third Xaa in SEQ ID NO:53 can be methionine (Met) or isoleucine (lie). The second Xaa in SEQ ID NO 59 can be Ala or Ser.

The antibody or antigen-binding fragment thereof may be an antibody or antigen-binding fragment thereof comprising: a heavy chain variable region comprising VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:33, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:39 and wherein Xaa 6 th and 7 th are Gly, and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:51, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:57 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63; or

An antibody or antigen-binding fragment thereof comprising: a heavy chain variable region comprising VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:38, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:44, and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 50; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:56, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:62 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 68.

The antibody or antigen-binding fragment thereof may be an antibody or antigen-binding fragment thereof comprising: a heavy chain variable region comprising a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:35 and wherein Xaa No. 2 is Tyr, a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:41 and wherein Xaa No. 8 is Ser, and a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO:47 and wherein Xaa No. 12 is His; and a light chain variable region comprising a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:53 and wherein Xaa No. 3 is Met, a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:59 and wherein Xaa No. 2 is Ala, and a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 65; or an antibody or antigen-binding fragment thereof comprising: a heavy chain variable region comprising VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:37, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:43 and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 49; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:55, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:61 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 67.

The antibody or antigen-binding fragment thereof may be an antibody or antigen-binding fragment thereof comprising: a heavy chain variable region comprising a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO. 33, a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO. 39 and wherein Xaa 6 and Xaa 7 are Ala and Gly, respectively, and a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO. 45; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:51, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:57 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63; or an antibody or antigen-binding fragment thereof comprising: a heavy chain variable region comprising VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:37, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:43 and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 49; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:55, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:61 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 67.

The antibody or antigen-binding fragment thereof may be an antibody or antigen-binding fragment thereof comprising: a heavy chain variable region comprising a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO. 36, a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO. 44 and a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO. 50; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:56, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:62 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 68; or an antibody or antigen-binding fragment thereof comprising: a heavy chain variable region comprising VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:37, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:43 and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 49; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:55, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:61 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 67.

The antibody or antigen-binding fragment thereof may comprise a heavy chain variable region comprising an amino acid sequence selected from any one of SEQ ID NOs 21 to 26; a light chain variable region comprising an amino acid sequence selected from any one of SEQ ID NOs 27 to 32; or the heavy chain variable region and the light chain variable region.

The antibody or antigen-binding fragment thereof may comprise the heavy chain variable region of SEQ ID NO 21 and the light chain variable region of SEQ ID NO 27; the heavy chain variable region of SEQ ID NO 22 and the light chain variable region of SEQ ID NO 28; the heavy chain variable region of SEQ ID NO. 23 and the light chain variable region of SEQ ID NO. 29; the heavy chain variable region of SEQ ID NO. 24 and the light chain variable region of SEQ ID NO. 30; the heavy chain variable region of SEQ ID NO. 25 and the light chain variable region of SEQ ID NO. 31; or the heavy chain variable region of SEQ ID NO 26 and the light chain variable region of SEQ ID NO 32; or a combination thereof.

Xaa 56 and 57 of SEQ ID NO. 21 can be Gly or Ala. In SEQ ID NO:23, Xaa 1 can be Glu (Gin) or Glu, Xaa 7 can be Ser or Pro, Xaa 12 can be Val or Ala, Xaa 27 can be Tyr or His, Xaa 58 can be Ser or Thr, Xaa 61 can be Asn or Ser, Xaa 74 can be Arg or Lys, Xaa 83 can be Phe (Phe) or Leu, Xaa 92 can be Gly or Ala, and Xaa 108 can be His or Tyr. The 53 rd Xaa in SEQ ID NO. 27 can be lie or Phe. In SEQ ID NO:29, Xaa 23 can be Met or lie, Xaa 45 can be Ala or Ser, and Xaa 73 can be Glu or aspartic acid (Asp), and Xaa 100 can be Met or lie.

The antibody or antigen binding fragment thereof may be a monoclonal antibody.

The antibody or antigen-binding fragment thereof may be labeled with a detectable label or a label capable of emitting a detectable signal. The label refers to a detectable compound or composition conjugated directly or indirectly to an antibody to produce a labeled antibody or antigen-binding fragment thereof. The label itself may be detectable and catalyze chemical modification of a detectable substrate compound or composition. The label may be an immunofluorescent label, a chemiluminescent label, a phosphorescent label, a radioactive label, an epitope tag, avidin/biotin, colloidal gold particles, colored particles, magnetic particles, chromophore labels, ECL labels, enzymes, and the like.

The antibody or antigen-binding fragment thereof can be produced by a hybridoma cell selected from the group consisting of the hybridoma cells deposited under accession numbers KCTC 137378P, KCTC 137388P, KCTC 137398P, KCTC 137408P, KCTC 137418P, KCTC 137428P, KCTC 137438P, KCTC 137448P, KCTC 137458P and KCTC 137468P.

The hybridoma cell refers to a hybrid cell having tumorigenicity generated by artificial fusion of two cells, and in general, can be used to continuously produce antibodies by fusing a B cell isolated from an immunized subject and a plasmacytoma cell. In the present specification, the hybridoma cell may be referred to as a hybridoma cell or a fused cell.

The cancer may be a solid or non-solid cancer. Solid cancer refers to the development of cancerous tumors in organs such as the liver, lungs, breast and skin. Non-solid cancers are cancers that occur in the blood and are also referred to as hematologic cancers. The cancer may be a carcinoma, sarcoma, hematopoietic-derived cancer, germ cell tumor, or blastoma. The cancer may be selected from, for example, breast cancer, colorectal cancer, head and neck cancer, colon cancer, skin cancer, pancreatic cancer, lung cancer, stomach cancer, prostate cancer, bladder cancer, urinary tract cancer, liver cancer, kidney cancer, clear cell sarcoma, melanoma, cerebrospinal tumor, brain cancer, thymus, mesothelioma, esophageal cancer, bile duct cancer, testicular cancer, germ cell tumor, thyroid cancer, parathyroid cancer, cervical cancer, endometrial cancer, lymphoma, myelodysplastic syndrome (MOS), myelofibrosis, acute leukemia, chronic leukemia, multiple myeloma, hodgkin's disease, endocrine cancer, and sarcoma.

The present inventors found that when using a combination of anti-BAG 2 antibodies or antigen-binding fragments thereof, in particular anti-BAG 2 antibodies or antigen-binding fragments thereof selected by BAG2 domain screening, the presence of BAG2 could be identified and its levels in cancer cell lines including breast, pancreatic, glioblastoma multiforme, gastric, ovarian or diffuse large B-cell lymphoma cell lines were significantly higher than in normal cells. Accordingly, the cancer diagnosed by using the composition may be selected from the group consisting of breast cancer, pancreatic cancer, glioma, gastric cancer, ovarian cancer, and lymphoma.

The present inventors found that when using an anti-BAG 2 antibody or antigen binding fragment thereof, and in particular a combination of an anti-BAG 2 antibody or antigen binding fragment thereof, BAG2 is overexpressed in breast cancer patients compared to normal subjects, and when BAG2 levels are high, the likelihood of a breast cancer patient suffering from metastatic breast cancer is high. Thus, the breast cancer diagnosed with the composition may be metastatic breast cancer.

Among the molecular subtypes of breast cancer, Triple Negative Breast Cancer (TNBC) is a very aggressive type with poor prognosis and high mortality despite systemic treatment. TNBC is heterogeneous compared to luminal or HER 2-enriched forms. While most targeted breast cancer therapies have shown positive results for hormone receptor and HER2 positive breast cancers, TNBC patients lack three types of target receptors including Estrogen Receptor (ER), Progestin Receptor (PR), human epidermal growth factor receptor (HER2), or other well-defined molecular targets, and therefore need to incorporate limited effective treatment options, e.g., poly ADP-ribose polymerase (PARP), Epidermal Growth Factor Receptor (EGFR), Src tyrosine kinase, etc. Therefore, to prevent unnecessary treatment routes and to select an effective treatment for TNBC patients, TNBC patients need to be quickly and accurately distinguished from various types of breast cancer patients.

The present inventors found that when using an anti-BAG 2 antibody or antigen-binding fragment thereof, and in particular a combination of an anti-BAG 2 antibody or antigen-binding fragment thereof, BAG2 is overexpressed in breast cancer patients, and when BAG2 levels are high, the probability that a breast cancer patient is diagnosed with TNBC-type breast cancer is high. Thus, the breast cancer that can be diagnosed by using the method may be triple negative breast cancer or TNBC type breast cancer.

The kit may further comprise one or more other component compositions, solutions or devices suitable for the analytical methods used in the kit, such as western blotting, ELISA, radioimmunoassay, radioimmunodiffusion, ouchterlony immunodiffusion, rocket immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, complement fixation, FAGS, protein chips, or combinations thereof. For example, in order to detect an immune complex of BAG2 in a sample using an antibody specific for each of them, the kit may further include a substrate, a suitable buffer, a second antibody labeled with a coloring enzyme or a fluorescent substance, or a colored substrate. The substrate may be a nitrocellulose membrane, a 96-well plate synthesized with polyethylene resin, a 96-well plate synthesized with polystyrene resin, a glass slide, or the like; the coloring enzyme may be peroxidase, alkaline phosphatase, etc.; the fluorescent substance may be Fluorescein Isothiocyanate (FITC), rhodamine B-isothiocyanate (RITC), etc.; and the coloring substrate may be 2,2' -azido-bis (3-ethylbenzothiazoline-6-sulfonic Acid) (ABTS), o-phenylenediamine (OPD), Tetramethylbenzidine (TMB), or the like.

The method is a method of providing information for diagnosing cancer, wherein the presence of BAG2 in a subject is detected to diagnose the presence of cancer in the subject.

The method may comprise contacting a sample isolated from a subject with an antibody that specifically binds to a BAG2 polypeptide or fragment thereof; and measuring the formed complex of BAG2 polypeptide or fragment thereof and antibody.

The method may further comprise determining whether the subject has cancer. The information may be information on whether the level of BAG2 measured in the subject is higher than, equal to, or lower than the level of BAG2 measured in the control group.

The sample may be a biological sample isolated from a subject to be diagnosed. The biological sample may be a cell, organ, cell lysate, whole blood, serum, plasma, lymph fluid, extracellular fluid, bodily fluid, urine, stool, tissue, bone marrow, saliva, sputum, cerebrospinal fluid, or a combination thereof.

The sample may be blood, serum, plasma, or a combination thereof. The present inventors have demonstrated that BAG2 is secreted extracellularly and that BAG2 is actually detected in the serum of breast cancer patients by using anti-BAG 2 antibodies or antigen-binding fragments thereof. Thus, BAG2 may be soluble in blood, serum, plasma, or combinations thereof.

When BAG2 is present in a sample isolated from a subject, an antibody that specifically binds to a BAG2 polypeptide or fragment thereof can bind to BAG2 in the sample. The antibody may be labeled, for example, with a fluorophore, a chromophore, or an enzyme capable of converting a substrate to a chromophore, e.g., to visualize the presence of BAG2 in a sample.

BAG2 in the sample forms a complex with an antibody that specifically binds to BAG2 polypeptide or fragment thereof as a result of the binding reaction, and from the complex, the presence or level of BAG2 can be identified or measured, if desired, using methods known to those skilled in the art. The complex can be measured by western blotting, enzyme-linked immunosorbent assay (ELISA), Radioimmunoassay (RIA), radioimmunodiffusion, ouchterlony immunodiffusion, rocket immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, complement fixation assay, FACS, protein chips, or a combination thereof. The measurement may be performed to measure the level of the complex in the sample.

The control group may be a sample taken from a healthy subject or a sample taken from breast cancer, pancreatic cancer, glioblastoma multiforme, gastric cancer, ovarian cancer or diffuse large B-cell lymphoma (DLBCL). Thus, the BAG2 level of the control group may be an average of BAG2 concentration in samples taken from healthy subjects, or, if desired, samples taken from breast cancer, pancreatic cancer, glioblastoma multiforme, gastric cancer, ovarian cancer, or DLBCL patients.

The sample used as a control may be of the same type as the sample taken at the same anatomical location for diagnosis. For example, when the sample is blood taken from the median cubital vein of the subject, the control may also be blood taken from the veins in the palate of the control group.

A healthy subject is a subject who does not have any acute or chronic disease, at least cancer, e.g., breast cancer, pancreatic cancer, glioblastoma multiforme, gastric cancer, ovarian cancer or DLBCL.

The level of BAG2 in a sample collected from a healthy subject may be substantially free of BAG 2. Thus, where the control group is set to have a value obtained from a healthy subject, the subject may be suspected of having cancer when the subject to be diagnosed is found to have BAG2, or a significantly high level of BAG2 therein. On the other hand, as shown in section 3 of example 2 of the present specification, in the blood of metastatic breast cancer patients, for example, in the blood of TNBC type patients among breast cancer patients, the possibility of identifying the presence of BAG2 was high, and the average value of BAG2 was significantly high (fig. 7 and 8). Thus, where the control group is set to a sample isolated from a breast cancer patient, a non-metastatic breast cancer patient or a non-TNBC type breast cancer patient, the subject may be suspected of having metastatic breast cancer or TNBC type breast cancer when the level of BAG2 is determined to be significantly high in the subject to be diagnosed.

Accordingly, in one aspect, the present invention relates to a method for detecting the presence of cancer in an individual, the method comprising contacting a biological sample isolated from the individual with an antibody or antigen-binding fragment thereof that specifically binds to a BAG2 polypeptide or fragment thereof; and measuring the formed complexes of the BAG2 polypeptide or fragment thereof and the antibody or antigen-binding fragment thereof, wherein the presence of cancer is detected if the level of BAG2 measured in the sample is higher than the level of BAG2 measured in the negative control group.

Drawings

This patent or application document contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the office upon request and payment of the necessary fee.

The present invention will become more fully understood from the detailed description given herein below and the accompanying drawings, which are given by way of illustration only, and thus are not limitative of the present invention, and wherein:

FIG. 1 shows the results of Western blotting of anti-BAG 2 antibody produced from 10 mouse hybridoma cells;

figures 2A and 2B show results of western blots of full length BAG2 polypeptide or fragment thereof of anti-BAG 2 antibody;

figure 3 shows the BAG2 domain reacting with a corresponding anti-BAG 2 antibody;

figure 4 shows a standard curve for BAG2 protein in possible 90 combinations of anti-BAG 2 antibodies;

FIG. 5 shows a graph of the specific binding capacity of selected 14 antibody combinations with respect to BAG2 protein;

FIG. 6 shows the differences in BAG2 expression patterns in various cancer cell lines observed using the antibody combination of 2A11-3F12 and the antibody combination of 9B12-3G 8;

figure 7 shows the significant difference in BAG2 protein expression in the serum of luminal and TNBC type breast cancer patients observed using the antibody combination of 2a11-3F12 and the antibody combination of 9B12-3G 8;

figure 8 shows the significant difference in BAG2 protein expression in the serum of luminal and TNBC type breast cancer patients observed using the antibody combination of 8C4-3G8 and the antibody combination of 10H7-3G 8; and is

Figures 9A to 9D show receiver operating characteristic curve (ROC) values and area under the curve (AUC) values for four antibody combinations.

Detailed Description

The following examples are provided for the purpose of illustrating the invention and are not to be construed as limiting.

Examples

Example 1 selection, sequencing and antigen-antibody reaction of anti-Bag 2 antibody

1. Selection of monoclonal antibodies targeting bag2 and amino acid sequence analysis thereof

The inventors selected antibodies targeting bag2, analyzed their amino acid sequences, and determined the Complementarity Determining Regions (CDRs) of each of the antibodies.

In detail, a gene encoding human BAG2 protein consisting of the nucleotide sequence of SEQ ID NO:70 consisting of the amino acid sequence of SEQ ID NO:69 was cloned into pCAGGS plasmid and linearized, and the linearized construct was then inoculated into the muscle of five 6-week-old female BALB/c mice by applying an electric shock. The constructs were inoculated intramuscularly 3 times at three week intervals and consisted of 100ug of DNA in 100ul of PBS. At this time, the plasmids of the control group were also treated in the same manner. In order to generate therapeutic and diagnostic antibodies, more effective DNA vaccine-based immunization strategies than protein-based antigen injection were performed. Blood was collected from the fundus vena cava or tail vein of the mice, and the blood was examined by an enzyme immunoassay showing serum antibody titer, and 3 days after the last immunization, the spleen was extracted from the mice showing sufficient antibody titer. The B lymphocytes were isolated from the spleen and then fused with myeloma cells cultured with the isolated B lymphocytes, namely, the SP2/0-Ag14 cell line of ATCC, to thereby obtain fused cells. After culturing the fused cells in HAT medium containing hypoxanthine, aminopterin and thymidine, hybridoma cells fused only with myeloma and B lymphocytes were obtained by selecting about 130 clones. Among the hybridoma cells obtained by the immunoblot selection process, 10 hybridoma cells producing an antibody specifically binding to human BAG2 protein were obtained.

Total RNA for anti-BAG 2 antibody was produced from 5x106 hybridoma cells and 5'-RACE-cDNA was generated from 100ng of total RNA by using SMART RACE cDNA amplification kit (Clontech) according to the manufacturer's instructions. The heavy chain variable region (VH) and light chain variable region (VL) coding regions were amplified by PCR, and the amplified genes were inserted into pGEM-T vector (Promega, USA), cloned, and the nucleotide sequences of the genes were analyzed by using an automatic gene analyzer (ABI Prism 310, Applied Biosystem Co.). The nucleotide sequence of the gene analyzed was identified by comparison with previously reported nucleotide sequences, and the identified nucleotide sequences were manually translated for determination of the sequences of complementarity determining regions VH-CDR1, -CDR2 and-CDR 3 and VL-CDR1, -CDR2 and-CDR 3. The determination of the sequence of the CDRs was performed using the Kabat database (http:// www.bioinf.org.uk/abs /).

As a result, 10 anti-BAG 2 antibodies that specifically bound to BAG2 were obtained from the hybridoma cells. The 10 anti-BAG 2 antibodies were the 2a11, 4C2, 8C4, 3B5, 9B3, 9B12, 3B10, 10H7, 3GB and 3F12 antibodies. In addition, the amino acid sequences of the heavy chain variable regions, the light chain variable regions and their complementarity determining regions shown in tables 1 to 3 and the nucleotide sequences of the genes encoding the antibodies were determined.

The 2A11, 4C2, and 8C4 antibodies comprised a heavy chain variable region consisting of the amino acid sequence of SEQ ID NO:21 and a light chain variable region consisting of the amino acid sequence of SEQ ID NO: 27. In the 2A11 antibody, 56 th and 57 th Xaa of SEQ ID NO:21 are each Gly, and 53 th Xaa of SEQ ID NO:27 is lie. In the 4C2 antibody, the 56 th Xaa and the 57 th Xaa of SEQ ID NO. 21 are Gly and Ala, respectively, and the 53 th Xaa of SEQ ID NO. 27 is Phe. In the 8C4 antibody, the 56 th Xaa and the 57 th Xaa of SEQ ID NO. 21 are Ala and Gly, respectively, and the 53 th Xaa of SEQ ID NO. 27 is Phe.

The VH-CDR1, -CDR2 and-CDR 3 of the 2A11, 4C2 and 8C4 antibodies consist of the amino acid sequences of SEQ ID NO:33, 39 and 45, respectively, and VL-CDR1, -CDR2 and-CDR 3 consist of the amino acid sequences of SEQ ID NO:51, 57 and 63, respectively. In the 2A11 antibody, Xaa 56 and 57 of SEQ ID NO 21 are each Gly. With respect to the 4C2 antibody, in SEQ ID NO:21, Xaa 56 is Gly and Xaa 57 is Ala. For the 8C4 antibody, in SEQ ID NO:21, Xaa 56 is Ala and Xaa 57 is Gly.

The 9B3, 9B12, and 3B10 antibodies comprised a heavy chain variable region consisting of the amino acid sequence of SEQ ID NO. 23 and a light chain variable region consisting of the amino acid sequence of SEQ ID NO. 29. With respect to the 9B3 antibody, in SEQ ID NO. 23, Xaa 1 is Glu, Xaa 7 is Ser, Xaa 12 is Val, Xaa 27 is Tyr, Xaa 58 is Ser, Xaa 61 is Asn, Xaa 74 is Lys, Xaa 83 is Phe, Xaa 92 is Ala, and Xaa 108 is Tyr. With respect to the 9B3 antibody, in SEQ ID NO:29, Xaa 23 is lie, Xaa 45 is Ala, Xaa 73 is Glu, and Xaa 100 is lie. With respect to the 9B12 antibody, in SEQ ID No. 23, Xaa 1 is Gin, Xaa 7 is Ser, Xaa 12 is Val, Xaa 27 is Tyr, Xaa 58 is Ser, Xaa 61 is Asn, Xaa 74 is Arg, Xaa 83 is Phe, Xaa 92 is Gly, and Xaa 108 is His. With respect to the 9B12 antibody, in SEQ ID NO:29, the 23 rd Xaa is Met, the 45 th Xaa is Ala, the 73 th Xaa is Glu, and the 100 th Xaa is Met. With respect to the 3B10 antibody, in SEQ ID No. 23, Xaa 1 is Gin, Xaa 7 is Pro, Xaa 12 is Ala, Xaa 27 is His, Xaa 58 is Thr, Xaa 61 is Ser, Xaa 74 is Arg, Xaa 83 is Leu, Xaa 92 is Gly, and Xaa 108 is His. With respect to the 3B10 antibody, in SEQ ID NO:29, Xaa 23 is Met, Xaa 45 is Ser, Xaa 73 is Asp, and Xaa 100 is lie.

With respect to the 9B3, 9B12, and 3B10 antibodies, VH-CDR1, -CDR2, and-CDR 3 consist of the amino acid sequences of SEQ ID NOS 35, 41, and 47, respectively, and VL-CDR1, -CDR2, and-CDR 3 consist of the amino acid sequences of SEQ ID NOS 53, 59, and 65, respectively. With respect to the 9B3 antibody, Xaa 2 of SEQ ID NO 35 is Tyr, Xaa 8 of SEQ ID NO 41 is Ser, Xaa 12 of SEQ ID NO 47 is Tyr, Xaa 3 of SEQ ID NO 53 is lie, and Xaa 2 of SEQ ID NO 59 is Ala. With respect to the 9B12 antibody, Xaa 2 of SEQ ID NO 35 is Tyr, Xaa 8 of SEQ ID NO 41 is Ser, Xaa 12 of SEQ ID NO 47 is His, Xaa 3 of SEQ ID NO 53 is Met, and Xaa 2 of SEQ ID NO 59 is Ala. With respect to the 3B10 antibody, Xaa No. 2 of SEQ ID NO:35 is His, Xaa No. 8 of SEQ ID NO:41 is Thr, Xaa No. 12 of SEQ ID NO:47 is His, Xaa No. 3 of SEQ ID NO:53 is Met, and Xaa No. 2 of SEQ ID NO:59 is Ser.

TABLE 1

Name of antibody	Nucleotide sequence of VH gene	Nucleotide sequence of VL gene
			2A11	SEQ ID NO:1	SEQ ID NO:11
4C2	SEQ ID NO:2	SEQ ID NO:12
			8C4	SEQ ID NO:3	SEQ ID NO:13
3B5	SEQ ID NO:4	SEQ ID NO:14
			9B3	SEQ ID NO:5	SEQ ID NO:15
9B12	SEQ ID NO:6	SEQ ID NO:16
			3B10	SEQ ID NO:7	SEQ ID NO:17
10H7	SEQ ID NO:8	SEQ ID NO:18
			3G8	SEQ ID NO:9	SEQ ID NO:19
3F12	SEQ ID NO:10	SEQ ID NO:20

TABLE 2

Name of antibody	Amino acid sequence of VH region	Amino acid sequence of VL region
			2A11	SEQ ID NO:21	SEQ ID NO:27
4C2	SEQ ID NO:21	SEQ ID NO:27
			8C4	SEQ ID NO:21	SEQ ID NO:27
3B5	SEQ ID NO:22	SEQ ID NO:28
			9B3	SEQ ID NO:23	SEQ ID NO:29
9B12	SEQ ID NO:23	SEQ ID NO:29
			3B10	SEQ ID NO:23	SEQ ID NO:29
10H7	SEQ ID NO:24	SEQ ID NO:30
			3G8	SEQ ID NO:25	SEQ ID NO:31
3F12	SEQ ID NO:26	SEQ ID NO:32

TABLE 3

2. Identification of antigen-antibody response of anti-BAG 2 antibody in breast cancer cells

Figure 1 shows the results of an immunoblot of anti-BAG 2 antibodies generated from 10 mouse hybridoma cells. Specifically, human breast cancer cells MDA-MB-231 cells were cultured in DMEM (Welgene) medium containing 10% FBS, 100U/ml penicillin and 100. mu.g/ml streptomycin at a temperature of 37 ℃. Cells were harvested from wells and washed with PBS, and the cells were lysed in lysis buffer containing 1% Brij 97, 5mM EDTA, 0.02M HEPES pH 7.3, 0.15M NaCl, 1mM PMSF, 0.5mM NaF, 10. mu.g/rni aprotinin, and 0.2mM sodium orthovanadate. After incubation on ice for 15 minutes, nuclei were removed from the cells by centrifugation and the supernatant was collected. To an appropriate amount of supernatant was added 2X sample buffer consisting of 20% glycerol, 4.6% SOS, 0.125M tris (pH 6.8), 0.1% bromophenol blue. SOS-PAGE analysis of 10ug protein samples was performed on 12% gels by using the mini-Protean II system (Bio-Rad Hercules, Calif.) under standard conditions. For immunoblotting, proteins were transferred to Millipore (a PVDF membrane). Blocking solutions consisting of 0.1

% Tween

20 and 5% Bovine Serum Albumin (BSA) in TBS were allowed to react for 1 hour. Subsequently, 1/2000 dilution of anti-BAG 2 antibody extracted from hybridoma cell culture was used as the primary antibody, and goat anti-mouse HRP conjugate (Dako) used as the secondary antibody was diluted to 1/5000. Film sensitization was performed in the dark using EGL reagent (Amersham Pharmacia Biotech) as substrate. The light sensitive bands were compared to standard molecular markers to identify bands corresponding to the size of BAG 2.

As a result, as shown in fig. 1, antibodies 2a11, 3B5, 3B10, 3F12, 3GB, 4C2, 8C4, 9B3, 9B12, and 10H7 exhibited antigen-antibody reactions targeting BAG2, compared to the commercially available polyclonal anti-BAG 2 antibody ab58682(Abcam) used as a positive control.

Next, in order to map the domains of BAG2 antigen against which ten antibodies identified in the above section 1 cells were directed, cells into which GST-empty vector (pcdna3.1+/GST vector, NovoPro Bioscience inc. GST-Bag complete vector, GST-Bag F1 vector, GST-Bag F2 vector, GST-Bag F3 vector, and GST-Bag F4 vector (each comprising a polynucleotide encoding a human BAG2 protein and a polynucleotide encoding a fragment of BAG2 protein) are introduced into cells, and the cells into which the vectors are introduced are cultured to express the genes, and then cell lysates are obtained. Immunoblotting was performed with each of the 10 antibodies against the cell lysate. The polynucleotide encoding human BAG2 protein has the nucleotide sequence of SEQ ID NO. 70. The GST-Bag F1, -Bag F2, -Bag F3 and-Bag F4 vectors consist of the nucleotide sequences of SEQ ID NOS 71 to 74, respectively.

Fig. 2A and 2B show the results of immunoblotting of a full-length BAG2 polypeptide of an anti-BAG 2 antibody, or a fragment thereof. In fig. 2, a shows a schematic diagram of the vector and BAG2 protein and fragments thereof, and B shows the results of immunoblotting. In detail, immunoblotting was performed as follows: each of the vectors was introduced into HEK293T cells by lipofectamine (lipofectamine) transfection (Thermo Fisher Scientific, inc., Waltham, MA, USA) method, and the resulting transformed cells were cultured in dmem (welgene) medium containing 10% FBS, 100U/ml penicillin and 100 μ g/ml streptomycin at 37 ℃ for 30 hours, and then the cells were isolated. The isolated cells were disrupted using the same method as described in connection with FIG. 1 and subjected to SOS-PAGE analysis on a 12% gel. For immunoblotting, after 1 hour of reaction with the same blocking solution as described in connection with fig. 1, each of 10 purified anti-BAG 2 antibodies at a concentration of 2mg/ml diluted in 1/10000 was used as a primary antibody and allowed to bind to cells. Goat anti-mouse HRP conjugate used as the secondary antibody was used at 1/5000 dilution concentration and film sensitization was performed in the dark using EGL reagent (Amersham Pharmacia Biotech) as substrate. Standard molecular marker sizes were expressed to confirm the size of BAG 2.

As a result, each anti-BAG 2 antibody differentially bound to a full-length BAG2 polypeptide or fragment thereof, as shown in figure 2. In particular, for each of the 10 anti-BAG 2 antibodies, a signal was typically detected at a position of about 50kDa in the cell lysate introduced with the GST-BAG intact vector. This result indicates that all of these antibodies can bind to full-length BAG2 polypeptide. Finally, the region of the BAG2 domain to which each anti-BAG 2 antibody reacts was identified.

Figure 3 shows the BAG2 domain reacted with a corresponding anti-BAG 2 antibody. As shown in fig. 3, 9B3, 9B12, 3B10, and 10H7 antibodies bound to the N-terminus of BAG2 protein, 2a11, 3B5, 4C2, and 8C4 antibodies bound to the middle region of BAG2 protein, and 3F12 and 3GB antibodies bound to the C-terminus of BAG2 protein. The N-terminus typically comprises a 21-60 amino acid coiled-coil region, and the middle region binds to a portion of the 109-189 amino acid BNB region. Thus, by using a set of antibodies that bind to different sites, BAG2 protein or a fragment thereof present in a sample can be detected with high sensitivity and specificity.

Example 2 screening of anti-BAG 2 antibody combination and confirmation of efficacy in cancer diagnosis

1. Combinations for screening anti-BAG 2 antibodies useful for cancer diagnosis

Figure 4 shows standard curves for BAG2 protein in possible 90 combinations of anti-BAG 2 antibodies. As a result, as shown in FIG. 4, among the possible combinations of anti-BAG 2 antibodies, 14 antibody combinations showing a standard curve with a high slope, namely 2A11-3F12, 10H7-3G8, 9B12-3G8, 10H7-3F12, 9B8-3G8, 3G8-3F12, 3B5-3F12, 3G8-4C2, 3F12-3G8, 3B5-3G8, 3B5-4C2, 3G8-8C4 and 9B3-3F12, were selected.

Figure 5 shows a graph of the specific binding capacity of selected 14 antibody combinations with respect to BAG2 protein. In detail, the Myc-tag BAG2 expression vector or Myc-tag empty vector was transduced into the human breast cancer cell line MDA-MB-231, respectively, by using Lipofectamine2000(lnvitrogen) according to the manufacturer's instructions. The relative amount of BAG2 protein secreted by 30ug lysate from control cells having Myc-tag empty vector introduced therein and cells overexpressing BAG2 protein having Myc-tag BAG2 expression vector introduced therein was determined using 14 antibody combinations. Control cells were labeled with EV and BAG2 overexpressing cells were labeled with OE. That is, by confirming the relative amounts of secreted BAG2 protein of BAG2 overexpressing cells and control cells, it was possible to determine the specific binding capacity of the antibody combination with respect to BAG2 protein.

As a result, as shown in FIG. 5, four antibody combinations having high specific binding ability to BAG2 protein, namely, 2A11-3F12, 9B12-3G8, 8C4-3G8 and 10H7-3F12 were selected.

2. Identification of the efficacy of selected anti-BAG 2 antibody combinations in the diagnosis of various cancers

To confirm the cancer diagnostic efficacy of the antibody combination selected in section 1, the antibody combination was confirmed to react with antigen-antibody of BAG 2. In detail, human BAG2 protein with 6 histidine attachments to the N-terminus produced and purified from human 293T cells was obtained as antigen.

Sandwich enzyme-linked immunosorbent assays (sandwich ELISA) were performed using each of the 2a11-3F12 and 9B12-3G8 antibody combinations.

In detail, sandwich ELISA was performed by the following procedure. Each of the antibodies 3F12 and 3G8 was reacted with 27ul of 10mM NHS-biotin (succinimidyl biotin, cat. No. 21435) in an amount of 1mg to prepare a biotin-bound detection antibody. Each of antibodies 2a11, 9B12, 8C4, and 10H7 as capture antibodies was diluted to a concentration of 3ug/ml using ELISA plate coating buffer (R & D system, DY006), and spread in an amount of 100ul in each well of a 96-well plate, and then coated overnight at a temperature of 4 ℃. The antibody-coated plates were blocked with 1% Bovine Serum Albumin (BSA) buffer (R & D system, DY995) for 1 hour at room temperature. Human His-tagged BAG2 recombinant protein (Ybiologics, Korea) produced and purified in the human-derived cell line HEK293T was prepared as BAG2 standard material at a concentration of 100ng/ml using 1% BSA buffer, and then diluted 4-fold to prepare standard solutions of 0.02, 0.097, 0.31, 1.56, 6.25 and 25 ng/ml. For the blood sample, 8ml or more of blood was collected from each of 14 healthy persons, 20 patients diagnosed with luminal breast cancer, and 38 patients diagnosed with TNBC type breast cancer, then mixed by gentle shaking, and left to stand at room temperature for 20 to 30 minutes. Then, the blood was centrifuged at 2,500rpm for 10 minutes. Sera stored below 4 ℃ were 1/2-fold diluted using 1% BSA buffer. 0, 0.02, 0.097, 0.39, 1.56, 6.25, 25 and 100ng/ml of BAG2 standard solution and 100ul of 1/2-fold diluted patient sample were dispensed into each well of the plate from which the blocking solution WAs removed and reacted at room temperature for 1 hour, followed by washing with washing buffer (R & D system, WA 126). 100ul of streptavidin-HRP (Pierce, 21130) diluted by 1:50000 in 1% BSA buffer was dispensed into each well and reacted at room temperature for 30 minutes, followed by washing with washing buffer. 100ul of 3,3',5,5' -tetramethylbenzidine (TMB, R & D system, DY999) was dispensed into each well, reacted at room temperature for 15 minutes in a cow, and then a stop solution was added thereto to stop the reaction. The concentration of BAG2 protein in the patient sample was calculated from a standard curve of the antibody obtained by measuring the absorbance of the reaction solution at 450 nm.

Fig. 6 shows the differences in expression patterns of BAG2 in cancer cell lines including breast cancer, pancreatic cancer, glioblastoma multiforme, gastric cancer, ovarian cancer and diffuse giant B-cell lymphoma, observed by using anti-BAG 2 antibody combinations. Fig. 6A shows the results of a sandwich ELISA with the 2a11-3F12 antibody combination, and fig. 6B shows the results of a sandwich ELISA with the 9B12-3G8 antibody combination.

As shown in fig. 6A and 6B, when the 2a11-3F12 and 9B12-3G8 antibodies were used in combination, the BAG protein was expressed at high levels in various cancer cell lines including breast cancer cell lines MDA-MB-231 and Hs578T, pancreatic cancer cell lines SNU2564 and PANC1, glioblastoma multiforme (GBM) cell lines U251MG and T98G, gastric cancer cell lines SNU1 and SNU484, ovarian cancer cell lines SKOV3 and a2780, and diffuse giant B-cell lymphoma (DLBCL) cell lines SU-DHL4 and SU-DHL 6. In contrast, BAG2 protein in normal cell lines T47D, SNU2469, a172, SNU620, OVCAR3 and U2932 was not expressed at all or expressed very little. In other words, it was confirmed by using 2a11-3F12 and 9B12-3G8 antibody in combination that BAG2 protein is significantly highly expressed in cancer cells including breast cancer, pancreatic cancer, glioblastoma multiforme, gastric cancer, ovarian cancer, and diffuse giant B-cell lymphoma, as compared to normal cells.

Therefore, when the above antibody combination is used, cancer can be effectively diagnosed by identifying whether the expression pattern of BAG2 protein is similar to that of BAG2 protein displayed in cancer cells, as compared to normal cells.

3. Identification of efficacy of selected anti-BAG 2 antibody combinations in diagnosing breast cancer

The differences in the expression pattern of BAG2 protein in serum between luminal and TNBC type breast cancer patients and normal subjects were confirmed by using the same sandwich ELISA method described in section 2. In detail, sera of healthy volunteers (N ═ 14), luminal breast cancer patients (N ═ 38), and TNBC type breast cancer patients (N ═ 38) were collected, and the expression pattern of BAG2 protein in the obtained sera was confirmed.

Figures 7A and 7B and 8A and 8B show significant differences in BAG2 protein expression in the serum of luminal and TNBG type breast cancer patients identified by using anti-BAG 2 antibody combinations. Fig. 7A shows the case of using the 9B12-3G8 antibody combination, and fig. 7B shows the case of using the 2a11-3F12 antibody combination. Fig. 8A shows the case of using the combination of 8G4-3G8 antibodies, and fig. 8B shows the case of using the combination of 10H7-3G8 antibodies.

As shown in fig. 7A and 7B and 8A and 8B, when the 9B12-3G8, 2a11-3F12, 8G4-3G8, and 1OH7-3F12 antibodies were used in combination, the p values for the respective antibody combinations were p <0.0001, p ═ 0.0373, p ═ 0.0009, p ═ 0.0190, indicating that there was a significant difference in the pattern of expression of BAG2 protein in the serum of luminal and TNBG-type breast cancer patients compared to normal subjects. Therefore, by observing significant differences in BAG2 protein expression patterns using antibody combinations, breast cancer can be effectively diagnosed.

4. The usefulness of selected anti-BAG 2 antibody combinations in breast cancer diagnosis was confirmed.

In order to confirm the sensitivity and specificity of cancer diagnosis using each of the four antibody combinations, the results of section 2 are shown by using the ROG curve.

Fig. 9A to 9D show recipient operational characteristics curves (ROG) and areas under curves (AUG) for the respective antibody combinations. Fig. 9A shows the case of using the combination of antibodies 9B12-3G8, fig. 9B shows the case of using the combination of antibodies 8G4-3G8, fig. 9C shows the case of using the combination of antibodies 2a11-3F12, and fig. 9D shows the case of using the combination of antibodies 10H7-3F 12.

As shown in fig. 9A to 9D, AUG values were 0.8596, 0.8368, 0.8554, and 0.6736 when combined using 9B12-3G8, 8G4-3G8, 2a11-3F12, and 10H7-3G8 antibodies. That is, the AUG value in the ROG curve was around 0.7 with all antibody combinations, and was between 0.8 and 0.9 when using the 9B12-3G8, 8G4-3G8, and 2a11-3F12 antibody combinations. These results indicate that each antibody combination can be used to diagnose breast cancer with high sensitivity and specificity. Therefore, the four antibody combinations can be effectively used for diagnosing breast cancer.

A composition for diagnosing cancer according to an aspect, comprising an anti-BAG 2 antibody or antigen-binding fragment thereof, may provide information for diagnosing cancer.

According to the method of providing information for diagnosing cancer according to another aspect, the presence or level of BAG2 polypeptide in blood can be identified or measured from blood, and the diagnosis is highly useful, unlike conventional diagnostic methods of collecting tissues. Thus, various organizations may use the methods to diagnose cancer.

[ accession number ]

The preservation organization: korea institute of bioscience and Biotechnology

Accession number: KCTC13737BP

The preservation date is as follows: 11/2018, 28/11/month

The preservation organization: institute of bioscience and biotechnology

Accession number: KCTC13738BP

The preservation date is as follows: 11/2018, 28/11/month

The preservation organization: korea institute of bioscience and Biotechnology

Accession number: KCTC13739BP

The preservation date is as follows: 11/2018, 28/11/month

The preservation organization: korea institute of bioscience and Biotechnology

Accession number: KCTC13740BP

The preservation date is as follows: 11/2018, 28/11/month

The preservation organization: korea institute of bioscience and Biotechnology

Accession number: KCTC13741BP

The preservation date is as follows: 11/2018, 28/11/month

The preservation organization: korea institute of bioscience and Biotechnology

Accession number: KCTC13742BP

The preservation date is as follows: 11/2018, 28/11/month

The preservation organization: korea institute of bioscience and Biotechnology

Accession number: KCTC13743BP

The preservation date is as follows: 11 and 28 months in 2018

The preservation organization: korea institute of bioscience and Biotechnology

Accession number: KCTC13744BP

The preservation date is as follows: 11/2018, 28/11/month

The preservation organization: institute of bioscience and biotechnology

Accession number: KCTC13745BP

The preservation date is as follows: 11 and 28 months in 2018

The preservation organization: institute of bioscience and biotechnology

Accession number: KCTC13746BP

The preservation date is as follows: 11/2018, 28/11/month

All references cited herein are incorporated by reference in their entirety.

*****

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention specifically described herein.

<110> McSt.McPad (Medpacto Inc.)

UNIVERSITY school property cooperative group of Chengyu UNIVERSITY (Research and Business Foundation sungkunnwan UNIVERSITY)

University of Yangshi educational Cooperation group (Industry-Academic Cooperation Foundation, Yonsei University)

<120> method for diagnosing cancer using anti-BAG 2 antibody

<130> 6987-0201

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<400> 9

cagatccagt tggtgcagtc tggacctgag ctgaagaagc ctggagagac agtcaagatc 60

tcctgcaagg cttctgggta tagcttcaca aagtatggaa tgaactgggt gaagcaggct 120

ccaggaaagg atatcaagtg gatggggtgg ataaacacca acactggaga ggcaacatat 180

ggtgaagagg tcaagggacg gtttgccttc tctttggaaa cctctgccag cactgcctat 240

ttgcagatca acaacctcaa aaatgaggac acggctacat atttctgtgc aagattggga 300

ttgaggtacc ttgactactg gggccaaggc accactctca cagtctcctc a 351

<210> 10

<211> 336

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3F12-VH

<400> 10

caggtgcaac tgcaggagtc aggacctgac ctggtgaaac cttctcagtc actttcactc 60

acctgcactg tcactgggta ctccatcacc agtggttata gctggcactg gatccggcag 120

tttccaggaa acaaattgga atggatgggc tacatatatt atagaggtag cactaactac 180

aacccatctc tcaaaagtcg aatctctatc actcgagaca catccaagaa ccagttcttc 240

ctgctgttga aatctgtgac tactgaggac acagccacat attactgtgc aagagaggct 300

tactggggcc aagggactct ggtcactgtc tcagca 336

<210> 11

<211> 336

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 2A11-VL

<400> 11

gatgttttga tgacccaaac tccactctcc ctgcctgtca gtcttggaga tcaagcctcc 60

atctcttgca gatctagtca gagtattgta catagtaatg gaaacaccta tttagaatgg 120

tacctgcaga agccaggcca gtctccaaag ctcctgatct acaaagtttc caaccgattt 180

tctggggtcc cagacaggtt cagtggcagt ggatcaggga cagatttcac actcaagatc 240

agcagagtgg aggctgagga tctgggagtt tatttctgct ttcaaggttc acaggttcct 300

ccgacgttcg gtggaggcac caagctggaa atcaaa 336

<210> 12

<211> 336

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 4C2-VL

<400> 12

gatgttttga tgacccaaac tccactctcc ctgcctgtca gtcttggaga tcaagcctcc 60

atctcttgca gatctagtca gagtattgta catagtaatg gaaacaccta tttagaatgg 120

tacctgcaga agccaggcca gtctccaaag ctcctgttct acaaagtttc caaccgattt 180

tctggggtcc cagacaggtt cagtggcagt ggatcaggga cagatttcac actcaagatc 240

agcagagtgg aggctgagga tctgggagtt tatttctgct ttcaaggttc acaggttcct 300

ccgacgttcg gtggaggcac caagctggaa atcaaa 336

<210> 13

<211> 336

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 8C4-VL

<400> 13

gatgttttga tgacccaaac tccactctcc ctgcctgtca gtcttggaga tcaagcctcc 60

atctcttgca gatctagtca gagtattgta catagtaatg gaaacaccta tttagaatgg 120

tacctgcaga agccaggcca gtctccaaag ctcctgttct acaaagtttc caaccgattt 180

tctggggtcc cagacaggtt cagtggcagt ggatcaggga cagatttcac actcaagatc 240

agcagagtgg aggctgagga tctgggagtt tatttctgct ttcaaggttc acaggttcct 300

ccgacgttcg gtggaggcac caagctggaa atcaaa 336

<210> 14

<211> 336

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3B5-VL

<400> 14

gatgttttga tgacccaaac tccactctcc ctgcctgtca gtcttggaga tcaagcctcc 60

atctcttgca gatccagtca gagccttgta cacagtaatg gaaacaccta tttacattgg 120

tacctgcaga agccaggcca gtctccaaag ctcctgatcc acaaagtttc caaccgattt 180

tctggggtcc cagacaggtt cagtggcagt ggatcaggga cagatttcac actcaagatc 240

agcagagtgg aggctgagga tctgggaatt tatttctgct ctcaaaatac acatattcct 300

ccgacgttcg ctggaggcac caagctggaa atcaaa 336

<210> 15

<211> 357

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 9B3-VL

<400> 15

gaggtccagc tgcaacaatc tggagctgag ctggtaaggc ctgggacttc agtgaaggtg 60

tcctgcaagg cttctggata cgccttcact aattacatga tagagtggat aaaacagagg 120

cctggacagg gccttgagtg gattggagtg attaatcctg gaagtggtgg tagttattac 180

aatgagaagg tcaagggcaa ggcaacactg accgcagaca aatcctccag cactgcctac 240

atgcagttca gcagcctgac agctgatgac tctgcggtct atttctgtcg gatctatggt 300

aactacaagg ggtactttga ctattggggc caaggcacca ctctcacagt ctcctca 357

<210> 16

<211> 321

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 9B12-VL

<400> 16

gacattgtga tgacccagtc tccatcttcc atgtatgcat ctctaggaga gagagtcact 60

atcacttgca aggcgagtca ggacatgaat agctatttaa gctggttcca gcagaaacca 120

gggaaatctc ctaagaccct gatctatcgt gcaaacagat tggtagatgg ggtcccatca 180

aggttcagtg gcagtggatc tgggcaagat tattctctca ccatcagcag cctggagtat 240

gaagatatgg gaatttatta ttgtctacag aatgatgagt ttccattcac gttcggctcg 300

gggacaaagc tggaaatgaa g 321

<210> 17

<211> 321

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3B10-VL

<400> 17

gacattgtga tgacccagtc tccatcttcc atgtatgcat ctctaggaga gagagtcact 60

atcacttgca aggcgagtca ggacatgaat agctatttaa gctggttcca gcagaaacca 120

gggaaatctc ctaagaccct gatctatcgt tcaaacagat tggtagatgg ggtcccatca 180

aggttcagtg gcagtggatc tgggcaagat tattctctca ccatcagcag cctggactat 240

gaagatatgg gaatttatta ttgtctacag aatgatgagt ttccattcac gttcggctcg 300

gggacaaagc tggaaataaa a 321

<210> 18

<211> 351

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 10H7-VL

<400> 18

gatgtccaga tgacccagtc tccatcctcc ctgtctgcat ctgtgggaga cagagccacc 60

atcacttgcc gggcaagtca gagcattagc agctatttaa attggtatca gcagaaaccc 120

gggaaagccc ctaagctcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180

aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag tctgcaacct 240

gaagattttg caacttacta ctgtcaacag agttacacta ccccgctcac tttcggtgga 300

ggcaccaagc tggaaatcaa acgtggagga gccagcctcg tggaattcaa g 351

<210> 19

<211> 333

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3G8-VL

<400> 19

gacattgtga tgacccagtc tcctgcttcc ttagttgtat ctctggggca gagggccacc 60

atctcatgca gggccagcaa aagtgtcagt acatctgact atagttatat gcactggtac 120

caacagaaac caggacagcc acccaaagtc ctcatctatc ttgcatccaa cctagaatct 180

ggggtccctg ccaggttcag tggcagtggg tctgggacag acttcaccct caacatccat 240

cctgtggagg aggaggatgc tgcaacctat tactgtcagc acaataggga gcttcctccc 300

acgttcggtg ctgggaccaa gctggagctg aaa 333

<210> 20

<211> 336

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3F12-VL

<400> 20

gatgttttga tgacccaaac tccactcact ttgtcggtta cctttgggca gccagcctcc 60

atctcttgca ggtcaagtca gagcctctta gatagtgatg gagagacata tttgaattgg 120

ttgttacaga ggccaggcca gtctccaaag cgcctaatct atctggtgtc taaactggac 180

tctggagtcc ctgacaggtt cactggcagt ggatcaggga cagatttcac actgaaaatc 240

agcagagtgg aggctgagga tttgggagtt tattattgct ggcaaggtac acattttccg 300

tacacgttcg gaggggggac caagctggaa ataaaa 336

<210> 21

<211> 123

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 2A11,4C2,8C4-VH

<220>

<221> variants

<222> (56)..(57)

<223> Xaa is Gly or Ala

<400> 21

Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg Pro Gly Ala

1 5 10 15

Ser Val Thr Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr

20 25 30

Glu Met His Trp Val Lys Gln Thr Pro Val His Gly Leu Glu Trp Ile

35 40 45

Gly Ile Ile Asp Pro Glu Thr Xaa Xaa Thr Ala Tyr Asn Gln Lys Phe

50 55 60

Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Gly Lys Phe Tyr Tyr Ser Gly Arg Asp Tyr Ala Met Asp Tyr

100 105 110

Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser

115 120

<210> 22

<211> 122

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3B5-VH

<400> 22

Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Asp Tyr

20 25 30

Thr Phe Tyr Trp Val Arg Gln Ser His Gly Glu Ser Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Asp Pro Tyr Asn Gly Gly Asn Thr Tyr Asn Arg Lys Phe

50 55 60

Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Phe

65 70 75 80

Met His Leu Asn Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Tyr Tyr Arg Tyr Gly Gly Gly Gly Asp Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Thr Leu Thr Val Ser Ser

115 120

<210> 23

<211> 119

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 9B3,9B12,3B10-VH

<220>

<221> variants

<222> (1)

<223> Xaa is Glu or Gln

<220>

<221> variants

<222> (7)

<223> Xaa is Ser or Pro

<220>

<221> variants

<222> (12)

<223> Xaa is Val or Ala

<220>

<221> variants

<222> (27)

<223> Xaa is Tyr or His

<220>

<221> variants

<222> (58)

<223> Xaa is Ser or Thr

<220>

<221> variants

<222> (61)

<223> Xaa is Asn or Ser

<220>

<221> variants

<222> (74)

<223> Xaa is Arg or Lys

<220>

<221> variants

<222> (83)

<223> Xaa is Phe or Leu

<220>

<221> variants

<222> (92)

<223> Xaa is Gly or Ala

<220>

<221> variants

<222> (108)

<223> Xaa is His or Tyr

<400> 23

Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Thr

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr

20 25 30

Met Ile Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Val Ile Asn Pro Gly Ser Gly Gly Ser Tyr Tyr Asn Glu Lys Val

50 55 60

Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Gln Phe Ser Ser Leu Thr Ala Asp Asp Ser Ala Val Tyr Phe Cys

85 90 95

Arg Ile Tyr Gly Asn Tyr Lys Gly Tyr Phe Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Thr Leu Thr Val Ser Ser

115

<210> 24

<211> 121

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 10H7-VH

<400> 24

Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu

1 5 10 15

Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr

20 25 30

Ser Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Phe Lys Trp Met

35 40 45

Gly Trp Ile Asn Thr Glu Thr Gly Glu Pro Thr Tyr Ala Asp Asp Phe

50 55 60

Lys Gly Arg Phe Ala Leu Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys

85 90 95

Ala Arg Phe Asp Tyr Gly Thr Ser Tyr Trp Tyr Phe Asp Val Trp Gly

100 105 110

Ala Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 25

<211> 117

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3G8-VH

<400> 25

Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu

1 5 10 15

Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Lys Tyr

20 25 30

Gly Met Asn Trp Val Lys Gln Ala Pro Gly Lys Asp Ile Lys Trp Met

35 40 45

Gly Trp Ile Asn Thr Asn Thr Gly Glu Ala Thr Tyr Gly Glu Glu Val

50 55 60

Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys

85 90 95

Ala Arg Leu Gly Leu Arg Tyr Leu Asp Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Leu Thr Val Ser Ser

115

<210> 26

<211> 112

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3F12-VH

<400> 26

Gln Val Gln Leu Gln Glu Ser Gly Pro Asp Leu Val Lys Pro Ser Gln

1 5 10 15

Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Gly

20 25 30

Tyr Ser Trp His Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp

35 40 45

Met Gly Tyr Ile Tyr Tyr Arg Gly Ser Thr Asn Tyr Asn Pro Ser Leu

50 55 60

Lys Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe Phe

65 70 75 80

Leu Leu Leu Lys Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg Glu Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala

100 105 110

<210> 27

<211> 112

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 2A11,4C2,8C4-VL

<220>

<221> variants

<222> (53)

<223> Xaa is Ile or Phe

<400> 27

Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly

1 5 10 15

Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser

20 25 30

Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Lys Leu Leu Xaa Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Phe Gln Gly

85 90 95

Ser Gln Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 28

<211> 112

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3B5-VL

<400> 28

Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly

1 5 10 15

Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser

20 25 30

Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Lys Leu Leu Ile His Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Phe Cys Ser Gln Asn

85 90 95

Thr His Ile Pro Pro Thr Phe Ala Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 29

<211> 107

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 9B3,9B12,3B10-VL

<220>

<221> variants

<222> (23)

<223> Xaa is Met or Ile

<220>

<221> variants

<222> (45)

<223> Xaa is Ala or Ser

<220>

<221> variants

<222> (73)

<223> Xaa is Glu or Asp

<220>

<221> variants

<222> (100)

<223> Xaa is Met or Ile

<400> 29

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Met Tyr Ala Ser Leu Gly

1 5 10 15

Glu Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Xaa Asn Ser Tyr

20 25 30

Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Lys Thr Leu Ile

35 40 45

Tyr Arg Xaa Asn Arg Leu Val Asp Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Gln Asp Tyr Ser Leu Thr Ile Ser Ser Leu Xaa Tyr

65 70 75 80

Glu Asp Met Gly Ile Tyr Tyr Cys Leu Gln Asn Asp Glu Phe Pro Phe

85 90 95

Thr Phe Gly Ser Gly Thr Lys Leu Glu Xaa Lys

100 105

<210> 30

<211> 107

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 10H7-VL

<400> 30

Asp Val Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Ala Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Thr Thr Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 31

<211> 111

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3G8-VL

<400> 31

Asp Ile Val Met Thr Gln Ser Pro Ala Ser Leu Val Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys Ser Val Ser Thr Ser

20 25 30

Asp Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Val Leu Ile Tyr Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His

65 70 75 80

Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gln His Asn Arg

85 90 95

Glu Leu Pro Pro Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys

100 105 110

<210> 32

<211> 112

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3F12-VL

<400> 32

Asp Val Leu Met Thr Gln Thr Pro Leu Thr Leu Ser Val Thr Phe Gly

1 5 10 15

Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Asp Gly Glu Thr Tyr Leu Asn Trp Leu Leu Gln Arg Pro Gly Gln Ser

35 40 45

Pro Lys Arg Leu Ile Tyr Leu Val Ser Lys Leu Asp Ser Gly Val Pro

50 55 60

Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Trp Gln Gly

85 90 95

Thr His Phe Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 33

<211> 8

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 2A11,4C2,8C4-VHCDR1

<400> 33

Gly Tyr Thr Phe Thr Asp Tyr Glu

1 5

<210> 34

<211> 8

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3B5-VHCDR1

<400> 34

Gly Tyr Ser Phe Thr Asp Tyr Thr

1 5

<210> 35

<211> 8

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 9B3,9B12,3B10-VHCDR1

<220>

<221> variants

<222> (2)

<223> Xaa is Tyr or His

<400> 35

Gly Xaa Ala Phe Thr Asn Tyr Met

1 5

<210> 36

<211> 8

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 10H7-VHCDR1

<400> 36

Gly Tyr Thr Phe Thr Asp Tyr Ser

1 5

<210> 37

<211> 8

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3G8-VHCDR1

<400> 37

Gly Tyr Ser Phe Thr Lys Tyr Gly

1 5

<210> 38

<211> 9

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3F12-VHCDR1

<400> 38

Gly Tyr Ser Ile Thr Ser Gly Tyr Ser

1 5

<210> 39

<211> 8

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 2A11,4C2,8C4-VHCDR2

<220>

<221> variants

<222> (6)..(7)

<223> Xaa is Gly or Ala

<400> 39

Ile Asp Pro Glu Thr Xaa Xaa Thr

1 5

<210> 40

<211> 8

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3B5-VHCDR2

<400> 40

Ile Asp Pro Tyr Asn Gly Gly Asn

1 5

<210> 41

<211> 8

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 9B3,9B12,3B5-VHCDR2

<220>

<221> variants

<222> (8)

<223> Xaa is Ser or Thr

<400> 41

Ile Asn Pro Gly Ser Gly Gly Xaa

1 5

<210> 42

<211> 8

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 10H7-VHCDR2

<400> 42

Ile Asn Thr Glu Thr Gly Glu Pro

1 5

<210> 43

<211> 8

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3G8-VHCDR2

<400> 43

Ile Asn Thr Asn Thr Gly Glu Ala

1 5

<210> 44

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3F12-VHCDR2

<400> 44

Ile Tyr Tyr Arg Gly Ser Thr

1 5

<210> 45

<211> 16

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 2A11,4C2,8C4-VHCDR3

<400> 45

Thr Arg Gly Lys Phe Tyr Tyr Ser Gly Arg Asp Tyr Ala Met Asp Tyr

1 5 10 15

<210> 46

<211> 15

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3B5-VHCDR3

<400> 46

Ala Arg Gly Tyr Tyr Arg Tyr Gly Gly Gly Gly Asp Phe Asp Tyr

1 5 10 15

<210> 47

<211> 12

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 9B3,9B12,3B10-VHCDR3

<220>

<221> variants

<222> (12)

<223> Xaa is Tyr or His

<400> 47

Arg Ile Tyr Gly Asn Tyr Lys Gly Tyr Phe Asp Xaa

1 5 10

<210> 48

<211> 14

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 10H7-VHCDR3

<400> 48

Ala Arg Phe Asp Tyr Gly Thr Ser Tyr Trp Tyr Phe Asp Val

1 5 10

<210> 49

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3G8-VHCDR3

<400> 49

Ala Arg Leu Gly Leu Arg Tyr Leu Asp Tyr

1 5 10

<210> 50

<211> 5

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3F12-VHCDR3

<400> 50

Ala Arg Glu Ala Tyr

1 5

<210> 51

<211> 11

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 2A11,4C2,8C4-VLCDR1

<400> 51

Gln Ser Ile Val His Ser Asn Gly Asn Thr Tyr

1 5 10

<210> 52

<211> 11

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3B5-VLCDR1

<400> 52

Gln Ser Leu Val His Ser Asn Gly Asn Thr Tyr

1 5 10

<210> 53

<211> 6

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 9B3,9B12,3B10-VLCDR1

<220>

<221> variants

<222> (3)

<223> Xaa is Ile or Met

<400> 53

Gln Asp Xaa Asn Ser Tyr

1 5

<210> 54

<211> 6

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 10H7-VLCDR1

<400> 54

Gln Ser Ile Ser Ser Tyr

1 5

<210> 55

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3G8-VLCDR1

<400> 55

Lys Ser Val Ser Thr Ser Asp Tyr Ser Tyr

1 5 10

<210> 56

<211> 11

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3F12-VLCDR1

<400> 56

Gln Ser Leu Leu Asp Ser Asp Gly Glu Thr Tyr

1 5 10

<210> 57

<211> 3

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 2A11,4C2,8C4-VLCDR2

<400> 57

Lys Val Ser

1

<210> 58

<211> 3

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3B5-VLCDR2

<400> 58

Lys Val Ser

1

<210> 59

<211> 3

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 9B3,9B12,3B10-VLCDR2

<220>

<221> variants

<222> (2)

<223> Xaa is Ala or Ser

<400> 59

Arg Xaa Asn

1

<210> 60

<211> 3

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 10H7-VLCDR2

<400> 60

Ala Ala Ser

1

<210> 61

<211> 3

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3G8-VLCDR2

<400> 61

Leu Ala Ser

1

<210> 62

<211> 3

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3F12-VLCDR2

<400> 62

Leu Val Ser

1

<210> 63

<211> 9

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 2A11,4C2,8C4-VLCDR3

<400> 63

Phe Gln Gly Ser Gln Val Pro Pro Thr

1 5

<210> 64

<211> 9

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3B5-VLCDR3

<400> 64

Ser Gln Asn Thr His Ile Pro Pro Thr

1 5

<210> 65

<211> 9

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 9B3,9B12,3B10-VLCDR3

<400> 65

Leu Gln Asn Asp Glu Phe Pro Phe Thr

1 5

<210> 66

<211> 9

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 10H7-VLCDR3

<400> 66

Gln Gln Ser Tyr Thr Thr Pro Leu Thr

1 5

<210> 67

<211> 9

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3G8-VLCDR3

<400> 67

Gln His Asn Arg Glu Leu Pro Pro Thr

1 5

<210> 68

<211> 9

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 3F12-VLCDR3

<400> 68

Trp Gln Gly Thr His Phe Pro Tyr Thr

1 5

<210> 69

<211> 211

<212> PRT

<213> Intelligent (Homo sapiens)

<400> 69

Met Ala Gln Ala Lys Ile Asn Ala Lys Ala Asn Glu Gly Arg Phe Cys

1 5 10 15

Arg Ser Ser Ser Met Ala Asp Arg Ser Ser Arg Leu Leu Glu Ser Leu

20 25 30

Asp Gln Leu Glu Leu Arg Val Glu Ala Leu Arg Glu Ala Ala Thr Ala

35 40 45

Val Glu Gln Glu Lys Glu Ile Leu Leu Glu Met Ile His Ser Ile Gln

50 55 60

Asn Ser Gln Asp Met Arg Gln Ile Ser Asp Gly Glu Arg Glu Glu Leu

65 70 75 80

Asn Leu Thr Ala Asn Arg Leu Met Gly Arg Thr Leu Thr Val Glu Val

85 90 95

Ser Val Glu Thr Ile Arg Asn Pro Gln Gln Gln Glu Ser Leu Lys His

100 105 110

Ala Thr Arg Ile Ile Asp Glu Val Val Asn Lys Phe Leu Asp Asp Leu

115 120 125

Gly Asn Ala Lys Ser His Leu Met Ser Leu Tyr Ser Ala Cys Ser Ser

130 135 140

Glu Val Pro His Gly Pro Val Asp Gln Lys Phe Gln Ser Ile Val Ile

145 150 155 160

Gly Cys Ala Leu Glu Asp Gln Lys Lys Ile Lys Arg Arg Leu Glu Thr

165 170 175

Leu Leu Arg Asn Ile Glu Asn Ser Asp Lys Ala Ile Lys Leu Leu Glu

180 185 190

His Ser Lys Gly Ala Gly Ser Lys Thr Leu Gln Gln Asn Ala Glu Ser

195 200 205

Arg Phe Asn

210

<210> 70

<211> 636

<212> DNA

<213> Intelligent (Homo sapiens)

<400> 70

atggctcagg cgaagatcaa cgctaaagcc aacgaggggc gcttctgccg ctcctcctcc 60

atggctgacc gctccagccg cctgctggag agcctggacc agctggagct cagggttgaa 120

gctttgagag aagcagcaac tgctgttgag caagagaaag aaatccttct ggaaatgatc 180

cacagtatcc aaaatagcca ggacatgagg cagatcagtg acggagaaag agaagaatta 240

aatctgactg caaaccgttt gatgggaaga actctcaccg ttgaagtgtc agtagaaaca 300

attagaaacc cccagcagca agaatcccta aagcatgcca caaggattat tgatgaggtg 360

gtcaataagt ttctggatga tttgggaaat gccaagagtc atttaatgtc gctctacagt 420

gcatgttcat ctgaggtgcc acatgggcca gttgatcaga agtttcaatc catagtaatt 480

ggctgtgctc ttgaagatca gaagaaaatt aagagaagat tagagactct gcttagaaat 540

attgaaaact ctgacaaggc catcaagcta ttagagcatt ctaaaggagc tggttccaaa 600

actctgcaac aaaatgctga aagcagattc aattag 636

<210> 71

<211> 270

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> GST-Bag F1 vector

<400> 71

atggctcagg cgaagatcaa cgctaaagcc aacgaggggc gcttctgccg ctcctcctcc 60

atggctgacc gctccagccg cctgctggag agcctggacc agctggagct cagggttgaa 120

gctttgagag aagcagcaac tgctgttgag caagagaaag aaatccttct ggaaatgatc 180

cacagtatcc aaaatagcca ggacatgagg cagatcagtg acggagaaag agaagaatta 240

aatctgactg caaaccgttt gatgggaaga 270

<210> 72

<211> 360

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> GST-Bag F2 vector

<400> 72

atggctcagg cgaagatcaa cgctaaagcc aacgaggggc gcttctgccg ctcctcctcc 60

atggctgacc gctccagccg cctgctggag agcctggacc agctggagct cagggttgaa 120

gctttgagag aagcagcaac tgctgttgag caagagaaag aaatccttct ggaaatgatc 180

cacagtatcc aaaatagcca ggacatgagg cagatcagtg acggagaaag agaagaatta 240

aatctgactg caaaccgttt gatgggaaga actctcaccg ttgaagtgtc agtagaaaca 300

attagaaacc cccagcagca agaatcccta aagcatgcca caaggattat tgatgaggtg 360

360

<210> 73

<211> 450

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> GST-Bag F3 vector

<400> 73

atggctcagg cgaagatcaa cgctaaagcc aacgaggggc gcttctgccg ctcctcctcc 60

atggctgacc gctccagccg cctgctggag agcctggacc agctggagct cagggttgaa 120

gctttgagag aagcagcaac tgctgttgag caagagaaag aaatccttct ggaaatgatc 180

cacagtatcc aaaatagcca ggacatgagg cagatcagtg acggagaaag agaagaatta 240

aatctgactg caaaccgttt gatgggaaga actctcaccg ttgaagtgtc agtagaaaca 300

attagaaacc cccagcagca agaatcccta aagcatgcca caaggattat tgatgaggtg 360

gtcaataagt ttctggatga tttgggaaat gccaagagtc atttaatgtc gctctacagt 420

gcatgttcat ctgaggtgcc acatgggcca 450

<210> 74

<211> 540

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> GST-Bag F4 vector

<400> 74

atggctcagg cgaagatcaa cgctaaagcc aacgaggggc gcttctgccg ctcctcctcc 60

atggctgacc gctccagccg cctgctggag agcctggacc agctggagct cagggttgaa 120

gctttgagag aagcagcaac tgctgttgag caagagaaag aaatccttct ggaaatgatc 180

cacagtatcc aaaatagcca ggacatgagg cagatcagtg acggagaaag agaagaatta 240

aatctgactg caaaccgttt gatgggaaga actctcaccg ttgaagtgtc agtagaaaca 300

attagaaacc cccagcagca agaatcccta aagcatgcca caaggattat tgatgaggtg 360

gtcaataagt ttctggatga tttgggaaat gccaagagtc atttaatgtc gctctacagt 420

gcatgttcat ctgaggtgcc acatgggcca gttgatcaga agtttcaatc catagtaatt 480

ggctgtgctc ttgaagatca gaagaaaatt aagagaagat tagagactct gcttagaaat 540

540

Claims

1. A composition for diagnosing cancer, the composition comprising: an antibody or antigen-binding fragment thereof that specifically binds to a BAG2 polypeptide or fragment thereof.

2. The composition of claim 1, wherein the antibody or antigen-binding fragment thereof comprises:

a heavy chain variable region comprising a complementarity determining region (VH-CDR)1 consisting of the amino acid sequence of SEQ ID NO:33, a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:39, and a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45; and a light chain variable region comprising a complementarity determining region (VL-CDR)1 consisting of the amino acid sequence of SEQ ID NO:51, a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:57, and a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO:63,

a heavy chain variable region comprising VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:34, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:40 and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 46; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO. 52, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO. 58 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO. 64,

a heavy chain variable region comprising VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:35, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:41 and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 47; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:53, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:59 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO:65,

a heavy chain variable region comprising VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:36, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:42 and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 48; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:54, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:60 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO:66,

a heavy chain variable region comprising VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:37, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:43 and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 49; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:55, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:61 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO:67, and

a heavy chain variable region comprising VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:38, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:44, and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 50; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:56, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:62 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 68.

3. The composition of claim 1, wherein the antibody or antigen-binding fragment thereof comprises:

a heavy chain variable region comprising an amino acid sequence selected from any one of SEQ ID NOs 21 to 26; and

a light chain variable region comprising any one amino acid sequence selected from the group consisting of SEQ ID NOs 27 to 32.

4. The composition of claim 1, wherein the antibody or antigen-binding fragment thereof comprises an antibody or antigen-binding fragment thereof comprising: a heavy chain variable region comprising VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:33, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:39 and wherein Xaa 6 and Xaa 7 are each Gly and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:51, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:57 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63; and

an antibody or antigen-binding fragment thereof comprising: a heavy chain variable region comprising a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO. 38, a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO. 44 and a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO. 50; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:56, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:62 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 68.

5. The composition of claim 1, wherein the antibody or antigen-binding fragment thereof comprises:

an antibody or antigen-binding fragment thereof comprising: a heavy chain variable region comprising a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:35 and wherein Xaa No. 2 is Tyr, a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:41 and wherein Xaa No. 8 is Ser, and a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO:47 and wherein Xaa No. 12 is His; and a light chain variable region comprising a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO. 53 and wherein Xaa at position 3 is Met, a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO. 59 and wherein Xaa at position 2 is Ala, and a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO. 65; and

an antibody or antigen-binding fragment thereof comprising: a heavy chain variable region comprising VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:37, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:43 and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 49; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:55, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:61 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 67.

6. The composition of claim 1, wherein the antibody or antigen-binding fragment thereof comprises an antibody or antigen-binding fragment thereof comprising: a heavy chain variable region comprising VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:33, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:39 and wherein Xaa 6 and Xaa 7 are Ala and Gly, respectively, and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:51, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:57 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63; and

7. The composition of claim 1, wherein the antibody or antigen-binding fragment thereof comprises an antibody or antigen-binding fragment thereof comprising: a heavy chain variable region comprising VH-CDR1 consisting of the amino acid sequence of SEQ ID NO:36, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:44 and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 50; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:56, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:62 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 68; and

8. The composition of claim 1, wherein the antibody or antigen-binding fragment thereof is a monoclonal antibody.

9. The composition of claim 1, wherein the antibody or antigen-binding fragment is labeled with a detectable label or a label capable of emitting a detectable signal.

10. The composition of claim 1, wherein the antibody or antigen-binding fragment thereof is produced by a hybridoma selected from the group consisting of the hybridoma deposited under accession numbers KCTC13737BP, KCTC 137388P, KCTC 137398P, KCTC13740BP, KCTC13741BP, KCTC13742BP, KCTC13743BP, KCTC13744BP, KCTC13745BP, and KCTC13746 BP.

11. The composition of claim 1, wherein the cancer is selected from, for example, breast cancer, colorectal cancer, head and neck cancer, colon cancer, skin cancer, pancreatic cancer, lung cancer, stomach cancer, prostate cancer, bladder cancer, urinary tract cancer, liver cancer, kidney cancer, clear cell sarcoma, melanoma, encephalomyelic tumor, brain cancer, thymus, mesothelioma, esophageal cancer, bile duct cancer, testicular cancer, germ cell tumor, thyroid cancer, parathyroid cancer, cervical cancer, endometrial cancer, lymphoma, myelodysplastic syndrome (MOS), myelofibrosis, acute leukemia, chronic leukemia, multiple myeloma, hodgkin's disease, endocrine cancer, and sarcoma.

12. A kit for diagnosing cancer, the kit comprising: the composition of any one of claims 1-11.

13. A method of providing information for diagnosing cancer in a subject, the method comprising: contacting a sample isolated from the subject with an antibody or antigen-binding fragment thereof that specifically binds to a BAG2 polypeptide or fragment thereof; and measuring the formed complex of the BAG2 polypeptide or fragment thereof and the antibody or antigen-binding fragment thereof.

14. The method of claim 13, wherein the antibody or antigen-binding fragment thereof comprises:

a heavy chain variable region comprising complementarity determining region (VH-CDR)1 consisting of the amino acid sequence of SEQ ID NO:33, VH-CDR2 consisting of the amino acid sequence of SEQ ID NO:39, and VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45; and a light chain variable region comprising a complementarity determining region (VL-CDR)1 consisting of the amino acid sequence of SEQ ID NO:51, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:57, and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO:63,

a heavy chain variable region comprising a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO. 34, a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO. 40 and a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO. 52, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO. 58 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO. 64,

a heavy chain variable region comprising a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO. 35, a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO. 41 and a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO. 47; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:53, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:59 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO:65,

a heavy chain variable region comprising a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO. 37, a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO. 43 and a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO. 49; and a light chain variable region comprising VL-CDR1 consisting of the amino acid sequence of SEQ ID NO:55, VL-CDR2 consisting of the amino acid sequence of SEQ ID NO:61 and VL-CDR3 consisting of the amino acid sequence of SEQ ID NO:67, and

15. The method of claim 13, wherein the sample is a biological sample.

16. The method of claim 15, wherein the biological sample is a cell, an organ, a cell lysate, whole blood, serum, plasma, lymph fluid, extracellular fluid, bodily fluid, urine, stool, tissue, bone marrow, saliva, sputum, cerebrospinal fluid, or a combination thereof.

17. The method of claim 13, wherein the cancer is selected from the group consisting of breast cancer, colorectal cancer, head and neck cancer, colon cancer, skin cancer, pancreatic cancer, lung cancer, stomach cancer, prostate cancer, bladder cancer, urinary tract cancer, liver cancer, kidney cancer, clear cell sarcoma, melanoma, encephalomyelial tumor, brain cancer, thymus, mesothelioma, esophageal cancer, cholangiocarcinoma, testicular cancer, germ cell tumor, thyroid cancer, parathyroid cancer, cervical cancer, endometrial cancer, lymphoma, myelodysplastic syndrome, myelofibrosis, acute leukemia, chronic leukemia, multiple myeloma, hodgkin's disease, endocrine cancer, and sarcoma.

18. The method of claim 13, wherein the information is information about whether the level of BAG2 measured in the subject is above, equal to, or below the level of BAG2 measured in a negative control group.

19. A method for detecting the presence of cancer in an individual, the method comprising

Contacting a biological sample isolated from the individual with an antibody or antigen-binding fragment thereof that specifically binds to a BAG2 polypeptide or fragment thereof; and

measuring the formed complexes of the BAG2 polypeptide or fragment thereof and the antibody or antigen-binding fragment thereof, wherein the presence of cancer is detected if the level of BAG2 measured in the sample is higher than the level of BAG2 measured in the negative control group.